Relation between activity-induced intracellular sodium transients and ATP dynamics in mouse hippocampal neurons

Abstract

Key points: Employing quantitative Na⁺ -imaging and Förster resonance energy transfer-based imaging with ATeam1.03^YEMK (ATeam), we studied the relation between activity-induced Na⁺ influx and intracellular ATP in CA1 pyramidal neurons of the mouse hippocampus. Calibration of ATeam in situ enabled a quantitative estimate of changes in intracellular ATP concentrations. Different paradigms of stimulation that induced global Na⁺ influx into the entire neuron resulted in decreases in [ATP] in the range of 0.1-0.6 mm in somata and dendrites, while Na⁺ influx that was locally restricted to parts of dendrites did not evoke a detectable change in dendritic [ATP]. Our data suggest that global Na⁺ transients require global cellular activation of the Na⁺ /K⁺ -ATPase resulting in a consumption of ATP that transiently overrides its production. For recovery from locally restricted Na⁺ influx, ATP production as well as fast intracellular diffusion of ATP and Na⁺ might prevent a local drop in [ATP].

Abstract: Excitatory neuronal activity results in the influx of Na⁺ through voltage- and ligand-gated channels. Recovery from accompanying increases in intracellular Na⁺ concentrations ([Na⁺ ]_i ) is mainly mediated by the Na⁺ /K⁺ -ATPase (NKA) and is one of the major energy-consuming processes in the brain. Here, we analysed the relation between different patterns of activity-induced [Na⁺ ]_i signalling and ATP in mouse hippocampal CA1 pyramidal neurons by Na⁺ imaging with sodium-binding benzofurane isophthalate (SBFI) and employing the genetically encoded nanosensor ATeam1.03^YEMK (ATeam). In situ calibrations demonstrated a sigmoidal dependence of the ATeam Förster resonance energy transfer ratio on the intracellular ATP concentration ([ATP]_i ) with an apparent K_D of 2.6 mm, indicating its suitability for [ATP]_i measurement. Induction of recurrent network activity resulted in global [Na⁺ ]_i oscillations with amplitudes of ∼10 mm, encompassing somata and dendrites. These were accompanied by a steady decline in [ATP]_i by 0.3-0.4 mm in both compartments. Global [Na⁺ ]_i transients, induced by afferent fibre stimulation or bath application of glutamate, caused delayed, transient decreases in [ATP]_i as well. Brief focal glutamate application that evoked transient local Na⁺ influx into a dendrite, however, did not result in a measurable reduction in [ATP]_i . Our results suggest that ATP consumption by the NKA following global [Na⁺ ]_i transients temporarily overrides its availability, causing a decrease in [ATP]_i . Locally restricted Na⁺ transients, however, do not result in detectable changes in local [ATP]_i , suggesting that ATP production, together with rapid intracellular diffusion of both ATP and Na⁺ from and to unstimulated neighbouring regions, counteracts a local energy shortage under these conditions.